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The functional characterization of an entire class of 17 genes from the red flour beetle,  Tribolium castaneum , which encode two families of  C uticular  P roteins  A nalogous to  P eritrophins (CPAPs) has been carried out.  CPAP  genes in  T. castaneum  are expressed exclusively in cuticle-forming tissues and have been classified into two families,  CPAP1  and  CPAP3 , based on whether the proteins contain either one (CPAP1), or three copies (CPAP3) of the chitin-binding domain, ChtBD2, with its six characteristically spaced cysteine residues. Individual members of the  TcCPAP1  and  TcCPAP3  gene families have distinct developmental patterns of expression. Many of these proteins serve essential and non-redundant functions in maintaining the structural integrity of the cuticle in different parts of the insect anatomy. Three genes of the  TcCPAP1  family and five genes of the  TcCPAP3  family are essential for insect development, molting, cuticle integrity, proper locomotion or fecundity. RNA interference (RNAi) targeting  TcCPAP1-C, TcCPAP1-H, TcCPAP1-J  or  TcCPAP3-C  transcripts resulted in death at the pharate adult stage of development. RNAi for  Tc C PAP3-A1, TcCPAP3-B, TcCPAP3-D1  or  TcCPAP3-D2  genes resulted in different developmental defects, including adult/embryonic mortality, abnormal elytra or hindwings, or an abnormal ‘stiff-jointed’ gait. These results provide experimental support for specialization in the functions of CPAP proteins in  T. castaneum  and a biological rationale for the conservation of CPAP orthologs in other orders of insects. This is the first comprehensive functional analysis of an entire class of cuticular proteins with one or more ChtBD2 domains in any insect species.

Gene Families of Cuticular Proteins Analogous to Peritrophins (CPAPs) in Tribolium castaneum Have Diverse Functions